Kit ɗin Methylation-specipc PCR (MSP)

Kit ɗin gano PCR na methylation.

Kit ɗin Methylation-musamman PC c PCR (MSP) an haɓaka shi musamman don abokan cinikin da ke nazarin halayen methylation na kwayoyin halittar DNA ta PCR, wanda MSP DNA Polymerase shine polymerase mai ƙarfi wanda aka gyara tare da ƙwayoyin rigakafi, kuma 10 × MSP PCR Buffer shine mai ba da PCR wanda aka inganta musamman don MSP dauki. Ya dace da TIANGEN DNA Bisulfite Kit ɗin Juyawa (4992447).

Cat. A'a Girman shiryawa
4992759 50 prep

Bayanin samfur

Gudun aiki

Misalan Gwaji

Tambayoyi

Alamar samfur

Siffofin

■ Samfurin yana da fa'idodin saurin sauri, sauƙi, babban hankali, takamaiman ƙarfi da kwanciyar hankali.

Musammantawa

Rubuta: Polymerase na DNA na MSP
Samfura: <500 na
Aikace -aikace: Ya dace da hanyar methylation takamaiman PCR (MSP) don nazarin halayen methylation na DNA genomic

Duk samfuran ana iya keɓance su don ODM/OEM. Don cikakkun bayanai,don Allah danna Sabis na Musamman (ODM/OEM)


  • Na baya:
  • Na gaba:

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    1. An yi amfani da Kit ɗin Methylation-Specific PCR (MSP) don haɓaka guntu 400 bp ta amfani da bisulfite da aka yi amfani da DNA na genomic azaman samfuri tare da tsarin amsawa na 20 μl.

    Experimental Exampl

    2. Saitin sake zagayowar PCR

    Experimental Exampl

     

    Experimental Examples 1: Samfuran da Mai sarrafa A ke sarrafa su;
    2: Samfuran da Mai Bayarwa B ke sarrafa su;
    3: samfurin TIANGEN da aka bi da shi 4: NTC; Saukewa: D2000
    Experimental Examples 1-6:
    6 maimaitawa;
    7: Mai Bayarwa Samfurin da aka sarrafa;
    8: NTC; Bio2-F/R da P16- Me-F/R: Nau'in gano abubuwa biyu.
    Tambaya: Babu makaɗan ƙarawa

    Samfurin A-1

    Template Samfurin yana ɗauke da ƙazamar furotin ko masu hana Taq, da dai sauransu ——- Tsarkake samfurin DNA, cire ƙazantar furotin ko cire samfuri na DNA tare da kayan tsarkakewa.

    ■ Ƙaddamar da samfuri bai cika ba ——Ya dace a ƙara yawan zazzabin denaturation da tsawaita lokacin denaturation.

    Deg ƙasƙantar da samfuri ——Ka sake shirya samfuri.

    Bayanan Bayani na A-2

    Quality Kyakkyawan ingancin fitila — —Re-synthesize primer.

    Deg ƙasƙantar da kai na farko — —Aliquot babban firam ɗin taro zuwa ƙaramin ƙara don adanawa. Guji daskarewa da narkewa da yawa ko adanawa na 4 ° C na dogon lokaci.

    Design Tsararrun ƙirar firamari (misali tsawon firamare bai isa ba, dimer da aka kafa tsakanin firam ɗin, da dai sauransu) -Rirar ƙira (ku guji samuwar dimer primer da tsarin sakandare)

    A-3 MG2+maida hankali

    ■ Mg2+ maida hankali yayi ƙasa sosai ——Yin haɓaka Mg daidai2+ maida hankali: Inganta Mg2+ maida hankali ta hanyar jerin halayen daga 1 mM zuwa 3 mM tare da tazara na 0.5 mM don ƙayyade mafi kyawun Mg2+ maida hankali ga kowane samfuri da share fage.

    A-4 Ƙara yawan zafin jiki

    Zazzabi mai ƙima yana shafar ɗaurin firamare da samfuri. —— Rage zafin zafin zafin jiki da inganta yanayin tare da ɗimbin digiri na 2 ° C.

    A-5 Tsawaita lokacin

    ■ Gajerun lokacin ƙara —— Ƙara lokacin ƙarawa.

    Tambaya: Tabbatacce

    Phenomena: Samfuran marasa kyau suma suna nuna makasudin jerin makasudin.

    A-1 Gurɓatar PCR

    Tsallake gurɓataccen jerin abubuwan da aka yi niyya ko samfuran ƙarfafawa ——Ayi hankali kada a ɗora samfurin da ke ɗauke da jerin manufa a cikin samfurin mara kyau ko zubar da su daga bututun ƙarfe. Yakamata reagents ko kayan aiki suyi aiki da kan su don kawar da acid nucleic da ke akwai, kuma yakamata a ƙaddara wanzuwar gurɓatarwa ta hanyar gwajin sarrafawa mara kyau.

    Contamin Gurɓatawa mai gurɓatawa ——Aliquot reagents da adanawa a ƙaramin zafin jiki.

    A-2 Firayim Ministar

    ■ Mg2+ maida hankali yayi ƙasa sosai ——Yin haɓaka Mg daidai2+ maida hankali: Inganta Mg2+ maida hankali ta hanyar jerin halayen daga 1 mM zuwa 3 mM tare da tazara na 0.5 mM don ƙayyade mafi kyawun Mg2+ maida hankali ga kowane samfuri da share fage.

    Tsarin ƙirar da bai dace ba, kuma jerin maƙasudin yana da homology tare da jerin marasa manufa. ——Re-design primers.

    Tambaya: Ƙarfafawa ba takamaimai ba

    Phenomena: Ƙungiyoyin haɓaka PCR ba su dace da girman da ake tsammani ba, ko babba ko ƙarami, ko kuma wani lokacin duka takamaiman ƙungiyoyin haɓakawa da ƙamus ɗin ba na musamman ba.

    Farashin A-1

    Ƙididdiga ta musamman

    ——Re-design primer.

    Concentration Mahimmancin taro ya yi yawa sosai ——Ya ƙara haɓaka zafin denaturation da tsawaita lokacin denaturation.

    A-2 MG2+ maida hankali

    M Mg2+ maida hankali ya yi yawa sosai ——Ya dace a rage maida hankali Mg2+: Inganta Mg2+ maida hankali ta hanyar jerin halayen daga 1 mM zuwa 3 mM tare da tazara na 0.5 mM don ƙayyade mafi kyawun Mg2+ maida hankali ga kowane samfuri da share fage.

    A-3 Polymerase mai ƙarfi

    Yawan adadin enzyme mai yawa ——Rage adadin enzyme yadda yakamata a tsakanin 0.5 U.

    A-4 Ƙara yawan zafin jiki

    Temperature Zazzabin ƙoshin ya yi ƙasa da yawa ——Ya dace a ƙara yawan zafin zafin ɗin ko a yi amfani da hanyar ƙara matakin biyu

    A-5 PCR hawan keke

    Cy Yawan hawan PCR da yawa —— Rage yawan hawan PCR.

    Tambaya: Ƙungiyoyi masu ƙyalli ko shafa

    Farashin A-1—— Bayanai marasa kyau ——Rayyana ƙirar fitila, canza matsayi da tsawon fitilar don haɓaka takamaiman sa; ko yin PCR da aka ƙera.

    A-2 Template DNA

    —— Samfurin ba shi da tsarki ——Ka tsarkake samfur ko cire DNA tare da kayan tsarkakewa.

    A-3 MG2+ maida hankali

    ——Mg2+ maida hankali yayi yawa ——Ya rage Mg daidai2+ maida hankali: Inganta Mg2+ maida hankali ta hanyar jerin halayen daga 1 mM zuwa 3 mM tare da tazara na 0.5 mM don ƙayyade mafi kyawun Mg2+ maida hankali ga kowane samfuri da share fage.

    A-4 dNTP

    ——Tinganin dNTPs sun yi yawa ——Rage taro na dNTP yadda yakamata

    A-5 Ƙara yawan zafin jiki

    —— Ƙaramin zafin zafin ƙanƙara mai zafi ——Ya dace a ƙara yawan zafin zafin

    Hanyoyin A-6

    ——Yawan hawan keke da yawa ——Ya inganta lambar zagayowar

    Tambaya: Nawa samfuri nawa ne ya kamata a ƙara a cikin tsarin amsawa na 50 μl PCR?
    ytry
    Tambaya: Yadda za a fadada dogayen gutsutsuren?

    Mataki na farko shine zaɓi polymerase da ya dace. Taq polymerase na yau da kullun ba zai iya sake karantawa ba saboda rashin aikin fitowar 3'-5 ', kuma rashin daidaituwa zai rage ƙimar fadada guntu. Sabili da haka, polymerase na Taq na yau da kullun ba zai iya haɓaka gutsattsarin da ya fi girma fiye da 5 kb ba. Taq polymerase tare da gyare -gyare na musamman ko wani babban aminci polymerase yakamata a zaɓi don haɓaka ingantaccen haɓakawa da saduwa da buƙatun ƙaramin guntu. Bugu da ƙari, ƙaramin guntun gutsutsuren kuma yana buƙatar daidaita daidaiton ƙirar fitila, lokacin denaturation, lokacin faɗaɗawa, pH mai ɓoyewa, da sauransu. Don hana lalacewar samfuri, lokacin denaturation a 94 ° C yakamata a rage zuwa 30 sec ko ƙasa da kowane zagayowar, kuma lokacin haɓaka zafin jiki zuwa 94 ° C kafin haɓaka ya zama ƙasa da 1 min. Haka kuma, saita zafin zafin a kusan 68 ° C da kuma tsara lokacin tsawaita gwargwadon ƙimar 1 kb/min na iya tabbatar da ingantaccen ingantaccen guntun gutsutsuren.

    Tambaya: Yadda za a inganta amincin amincin PCR?

    Ana iya rage ƙimar kuskuren haɓaka PCR ta amfani da polymerases DNA daban -daban tare da babban aminci. Daga cikin duk polymerases na Taq DNA da aka samo zuwa yanzu, enfuz enzyme yana da mafi ƙarancin ƙimar kuskure da mafi aminci (duba tebur da aka makala). Baya ga zaɓin enzyme, masu bincike na iya ƙara rage ƙimar maye gurbi ta PCR ta hanyar inganta yanayin amsawa, gami da haɓaka abun da ke ciki, maida hankali na polymerase mai ɗorewa da haɓaka lambar sake zagayowar PCR.

    Rubuta saƙonka a nan ka aika mana